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通过 EST 和微阵列策略进行花生感染黄曲霉的基因表达谱分析和抗性基因鉴定。

Gene expression profiling and identification of resistance genes to Aspergillus flavus infection in peanut through EST and microarray strategies.

机构信息

Crop Protection and Management Research Unit, U.S. Department of Agriculture-Agricultural Research Service, Tifton, GA, USA.

出版信息

Toxins (Basel). 2011 Jul;3(7):737-53. doi: 10.3390/toxins3070737. Epub 2011 Jun 24.

Abstract

Aspergillus flavus and A. parasiticus infect peanut seeds and produce aflatoxins, which are associated with various diseases in domestic animals and humans throughout the world. The most cost-effective strategy to minimize aflatoxin contamination involves the development of peanut cultivars that are resistant to fungal infection and/or aflatoxin production. To identify peanut Aspergillus-interactive and peanut Aspergillus-resistance genes, we carried out a large scale peanut Expressed Sequence Tag (EST) project which we used to construct a peanut glass slide oligonucleotide microarray. The fabricated microarray represents over 40% of the protein coding genes in the peanut genome. For expression profiling, resistant and susceptible peanut cultivars were infected with a mixture of Aspergillusflavus and parasiticus spores. The subsequent microarray analysis identified 62 genes in resistant cultivars that were up-expressed in response to Aspergillus infection. In addition, we identified 22 putative Aspergillus-resistance genes that were constitutively up-expressed in the resistant cultivar in comparison to the susceptible cultivar. Some of these genes were homologous to peanut, corn, and soybean genes that were previously shown to confer resistance to fungal infection. This study is a first step towards a comprehensive genome-scale platform for developing Aspergillus-resistant peanut cultivars through targeted marker-assisted breeding and genetic engineering.

摘要

黄曲霉和寄生曲霉感染花生种子并产生黄曲霉毒素,这种毒素与世界各地的家畜和人类的各种疾病有关。降低黄曲霉毒素污染最具成本效益的策略是开发抗真菌感染和/或抗黄曲霉毒素产生的花生品种。为了鉴定与花生中曲霉相互作用和花生抗曲霉的基因,我们进行了大规模的花生表达序列标签(EST)项目,我们用该项目来构建花生玻片寡核苷酸微阵列。所构建的微阵列代表了花生基因组中 40%以上的编码蛋白基因。为了进行表达谱分析,将抗感品种的花生与黄曲霉和寄生曲霉孢子的混合物进行感染。随后的微阵列分析鉴定出在抗感品种中,有 62 个基因在曲霉感染时呈上调表达。此外,我们还鉴定出 22 个假定的抗曲霉基因,这些基因在抗感品种中均呈组成性上调表达。其中一些基因与先前被证明可抵抗真菌感染的花生、玉米和大豆基因具有同源性。本研究是通过靶向标记辅助选择和遗传工程开发抗曲霉的花生品种的全面基因组规模平台的第一步。

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